原花青素对玉米赤霉烯酮诱导小鼠睾丸支持细胞氧化损伤的保护作用

发布时间：2018-04-17 08:00

  本文选题：原花青素 + 玉米赤霉烯酮　； 参考：《沈阳农业大学》2017年硕士论文

【摘要】：背景:玉米赤霉烯酮(ZEA)又叫作F-2毒素,是一种由镰孢菌属真菌产生的具有雌激素活性的非甾醇类真菌毒素,与体内雌二醇受体具有特殊亲和力。ZEA作为一种生物毒素具有特殊的毒性,其毒性作用主要表现在它的强雌激素样作用,直接对动物机体生殖系统造成严重损伤,造成动物繁殖功能下降,对畜牧业造成严重经济损失。原花青素(PC)作为一种国际上公认的最有效的清除人体内自由基的天然抗氧化剂具有极强的抗氧化作用,能有效清除超氧阴离子自由基和羟基自由基,且其来源十分广泛,本试验中选用PC作为ZEA氧化损伤作用的抗氧化剂来研究PC是否可以有效拮抗ZEA对鼠睾丸支持细胞的氧化损伤。目的:本试验旨在研究体外实验中PC在ZEA诱导的鼠睾丸支持细胞氧化损伤中的保护作用及其作用机制,为畜牧业生产中ZEA酮中毒的防治提供理论依据。方法:利用MTT法分别测出不同浓度的ZEA和PC单独和联合对鼠睾丸支持细胞相对存活率;流式细胞术法检测各组支持细胞凋亡率;利用试剂盒检测各组细胞T-SOD、LDH、GSH-Px活性及MDA和GSH含量;RT-PCR法分别检测核因子E2相关因子2(Nrf2)、血红素加氧酶1(HO-1)、谷胱甘肽过氧化物酶(GSH-Px)、醌氧化还原酶1(NQO1)、γ-谷氨酰半胱氨酸合成酶(γ-GCS)的mRNA表达水平;利用Western-blot法分别检测各组细胞Nrf2、GSH-Px、HO-1、γ-GCS、NQO1的蛋白表达水平。结果:1、ZEA对支持细胞具有损伤作用,低浓度的PC对支持细胞有促进生长的作用,PC对经ZEA染毒的支持细胞具有保护作用。2、与ZEA染毒组相比,PC能够降低经ZEA染毒的支持细胞的凋亡率(p0.01);与ZEA染毒组相比,PC能够提高T-SOD、GSH-PX活性(p0.05,p0.05),降低MDA含量,显著降低LDH活性(p0.05),显著提高GSH含量(p0.05)。3、与ZEA组相比,PC能够显著上调生精细胞Nrf2及其下游靶基因GSH-Px、HO-1、γ-GCS、NQO1的mRNA和蛋白表达量。结论:PC对ZEA诱导的鼠睾丸支持细胞氧化损伤具有保护作用,PC对ZEA诱导的鼠睾丸支持细胞损伤的保护作用可能是通过激活Nrf2/ARE通路途径实现。
[Abstract]:Background: ZEAA, also known as F-2 toxin, is an estrogen-active nonsterol mycotoxin produced by Fusarium fungi.As a biotoxin, ZEA has a special toxicity. Its toxic effect is mainly due to its strong estrogen-like effect, which directly causes serious damage to the reproductive system of animals.Causes the animal reproduction function to decline, causes the serious economic loss to the animal husbandry.As one of the most effective natural antioxidants for scavenging free radicals in human body, proanthocyanidin (PCP) has a strong antioxidant effect, and can effectively scavenge superoxide anion radical and hydroxyl radical, and its sources are very extensive.In this study, PC was used as an antioxidant for oxidative damage of ZEA to investigate whether PC could effectively antagonize oxidative damage of rat testicular Sertoli cells induced by ZEA.Objective: to study the protective effect and mechanism of PC on oxidative injury of testis Sertoli cells induced by ZEA in vitro, and to provide theoretical basis for the prevention and treatment of ZEA ketone poisoning in animal husbandry.Methods: the relative survival rate of rat testicular Sertoli cells with different concentrations of ZEA and PC was determined by MTT method, and the apoptosis rate of Sertoli cells in each group was detected by flow cytometry.The activity of T-SODX LDH-Px and the contents of MDA and GSH were detected by RT-PCR, and the mRNA expression levels of nuclear factor E2 related factor 2Nrf2, heme oxygenase 1HO-1, glutathione peroxidase (GSH-PxX), quinone redox enzyme 1NQO1, 纬 -glutamyl cysteine synthase (纬 -GCSA) were detected by RT-PCR.Western-blot assay was used to detect the protein expression level of Nrf2GCSNQO1, GSH-PxHO-1 and 纬 -GCSO-1 in each group.Results [WT5HZ] ZEA could injure Sertoli cells.Low concentration of PC can promote the growth of Sertoli cells. PC has protective effect on Sertoli cells exposed to ZEA. Compared with ZEA group, it can reduce the apoptosis rate of Sertoli cells exposed to ZEA, and compared with ZEA group, it can reduce the apoptosis rate of Sertoli cells.The activity of GSH-PX in T-SODX was increased, and the activity of GSH-PX was increased. The activity of GSH-PX was increased, and the content of MDA was decreased.Compared with ZEA group, PPC could significantly up-regulate the expression of mRNA and protein in spermatogenic cell Nrf2 and its downstream target genes GSH-PxHO-1, 纬 -GCSNQO1.Conclusion the protective effect of ZEA on oxidative injury of rat testicular Sertoli cells induced by ZEA may be realized by activating the pathway of Nrf2/ARE pathway in ZEA induced rat testicular Sertoli cells injury.
【学位授予单位】：沈阳农业大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S859.87
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本文编号：1762749